Novel steroid ester

ABSTRACT

21-M-SULFOBENZOATES OF 6,16 Alpha -DIMETHYL-2&#39;&#39;-PHENYL- Delta 2,4,6-PREGNATRIENEOLO (3,2-C)-PYRAZOL-11 Beta , 17, 21-TRIOL-20ONE OF THE FORMULA   WHEREIN R is selected from the group consisting of hydrogen and alkali metal useful as anti-inflammatory agents having prolonged activity and their preparation.

United States Patent [1 1 Pierdet I Mar. 25, I975 NOVEL STEROID ESTER[75] Inventor: Andre Pierdet, Noisy-le-Sec, France [73] Assignee:Roussel Uclaf, Paris, France [22] Filed: Dec. 11, 1973 [21] Appl. No.:423,663

Related U.S. Application Data [62] Division of Ser. No. 330,407, Feb. 7,1973, Pat. No.

[30] Foreign Application Priority Data Feb. 18, 1972 France 72.05527[52] US. Cl. 424/241 [51] Int. Cl A61k 17/00 [58] Field of Search424/241 [56] References Cited UNITED STATES PATENTS 3,037,034 5/1962Jolz et al 260/397.45 3,657,434 4/1972 Radscheit et al. 424/241 PrimaryE.\'aminerl-lenry A. French Attorney, Agent, or Firm-Hammond & Littell 57] ABSTRACT 2l-m-sulfobenzoates of 6,16a-dimethyl-2-phenyl-A-pregnatrieneolo [3,2-cl-pyrazol-l 1B, 17, Zl-triol- ZO-one of theformula wherein R is selected from the group consisting of hydrogen andalkali metal useful as anti-inflammatory agents having prolongedactivity and their preparation.

4 Claims, No Drawings NOVEL STEROID ESTER This is a division of Ser. No.330,407, filed Feb. 7, 1973 now U.S. Pat. No. 3,803,132.

STATE OF THE ART French Pat. No. 1,482,808 describes the alcohol, 6,l6B-dimethyl-2-phenyl-A2.4,6-pregnatrieneolo [3,2- c]-pyrazol-l 1B. 17,2l-triol-20-one and the simple esters thereof such as the acetate esterand their antiinflammatory activity.

OBJECTS OF THE INVENTION It is an object of the invention to provide thenovel 2l-m-sulfobenzoates of formula I.

It is another object of the invention to provide a novel process for thepreparation of the 21'msulfobenzoates of formula I.

It is an additional object of the invention to provide novelanti-inflammatory compositions having pro longed activity.

It is a further object of the invention to provide a novel method ofrelieving inflammation in warmblooded animals.

These and other objects and advantages of the invention will becomeobvious from the following detailed description.

THE INVENTION esters of the invention are 21-m- 16a-dimethyl-2'-phenyl-A"*"- l7, 2l-triol- The novel sulfobenzoates of 6,pregnatrieneolo [3,2-c]-pyrazol-l 1,8,

20-one of the formula wherein R is selected from the group consisting ofhydrogen and alkali metal. These esters have a considerably moreprolonged anti-inflammatory activity than theknown esters thereofdescribed in French Pat. No. 1,482,808.

The novel process for the preparation of the 21-msulfobenzoates offormula I comprises reacting the steroid alcohol of the formula with anesterifying derivative of methane sulfonic acid to obtain thecorresponding 2l-mesylate, reacting the latter with a double salt ofm-sulfobenzoic acid in the presence of a dialkyl carboxylic acid amideand passing the resulting product through an ion exchange resin in theacid form to obtain the product of formula I wherein R is hydrogen. Ifdesired, the latter may be re' acted with an alkali metal hydroxide toobtain the corresponding alkali metal salt of formula I.

In a preferred embodiment of the process, the 21- mesylate is reacted atabout C with disodium msulfobenzoate in the presence ofdimethylformamide and passing the product obtained through an ion exchange resin in the acid form to form the product of formula I wherein Ris hydrogen. If desired, the latter may be reacted with sodium hydroxideto form the corresponding sodium salt although other alkali metalhydroxides such as lithium or potassium hydroxide.

The novel anti-inflammatory compositions having prolonged activity ofthe invention are comprised of an effective amount of a compound offormula I and a pharmaceutical carrier. The said compositions may be inthe form of simple tablets, dragees, gelules, granules, solutionssuspensions, syrups, suppositories, pomades, cremes, gels and aerosolsand in the form of injectable solutions or suspensions or sterilepowders for extemporaneous dissolution made by the usual methods.

The excipient used for the pharmaceutical carrier may be any of thoseusually used such as talc, arabic gum, lactose, amidon, magnesiumstearate, cacao but ter, aqueous or non-aqueous vehicles, fattysubstances of animal or vegetable origin, paraffin derivatives, glycols,diverse wetting agents, dispersants or emulsifiers and preservatives.

The compositions of the invention due to their remarkableanti-inflammatory activity are very useful in therapy, for example, inhumans for the treatment of acute or chronic rheumatism, inflammatorydermatosis, asthma and viral hepatitis. Clinical experimentation hasconfirmed the remarkable activity of the products for the treatment ofasthma crisis. The regular and prw longed action of the compound offormula I wherein R is hydrogen has been ascertained in which certainknown derivatives where slightly active.

EXAMPLE I STEP A: 2l-mesylate of 6,loa-dimethyl-T-phenyl- A-pregnatrieneolo [3,2-c1-pyrazol-l lB,17,2l-triol- 20-one A solution of10.33 g (0.0212 mole) of 6,16a-. dimethyl-2-pheny1-A -pregnatrieneolo[3,2-c1- pyrazol-l 1B,17,21-triol-20-one in 60 ml of methylethylpyridinewas stirred for 10 minutes at 24C and 3.3 ml (0.0424 mole) of methanesulfonyl chloride was added to the resulting solution over minutes. Thetemperature was raised from 24C to 28C over minutes and the mixture wascooled on an ice bath to 10 i 2C which temperature was maintained for 1hour. The resulting mixture was added to 250 ml of distilled water and40 ml of concentrated hydrochloric acid and the mixture was stirred fortwo hours and then the pH was verified to be 1. The mixture wasextracted with ethyl acetate and the organic phase was washed with waterto a pH of 4-5, was dried over sodium sulfate and evaporated to drynessunder reduced pressure to obtain an amorphous product. The latter waschromatographed over silica gel eluting with a 4-6 ethyl acetatebenzenemixture to obtain 7.7 g of a white amorphous product which wascrystallized from l-l acetonewater mixture. After vacuum filtration, theproduct was washed and dried to obtain 6.56 g (55% yield) of 21-mesylate of 6,16adimethyl-2'-phenyl-A pregnatrieneolo [3,2-c]-pyrazol l1,8,17,21-triol- 20-one melting at 195C and having a specific rotation[11],, l.5: :1(c 0.64% in chloroform). The product occurred as whitecrystals soluble in ethanol, chloroform, acetone and ethyl acetate andinsoluble in water. STEP B: 2phenyl-A -pregnatrieneolo 11,8,17,21-triol-20-one 1.03 g (0.0123 mole) of sodium bicarbonate wereadded to a mixture of 3 g (0.0132 mole) of monoso dium m-sulfobenzoatein 3 ml of distilled water heated to 90 95C and after stirring for 10minutes, the mixture was added to 100 ml of dimethylformamide. Themixture was distilled to entrain all the water and was cooled to 95C.Then, 5 g (0.0088 mole) ofthe product ofStep A was added thereto all atonce and the mixture was stirred at 95C for 5 hours under a current ofnitrogen. The mixture was concentrated to dryness under reduced pressureand the residue was taken up in 100 ml of a solution of ethanolcontaining 50% water.

The resulting solution was passed through an ion exchange resin(Dowex 50type) in the acid form. The eluate was recovered and the alcohol wasdistilled off under a reduced pressure of 12 mm Hg and a temperature50C. An acid crystallized and was recovered by vacuum filtration and waswashed 4 times with distilled water. After a second passage through theion exchange resin followed by a distillation under reduced pressure, acreme solid product was obtained which was dissolved in 60 ml ofacetone. 60 ml of water were added thereto and the solution wasconcentrated. The crystals formed were recovered by vacuum filtrationand were washed with a 15-60 acetone-water mixture. The product wasredissolved in 70 ml of acetone and after 60 ml of water were added, thesolution was concentrated. The crystals formed were recovered by vacuumfiltration and were washed with a 25-60 acetonewater mixture. The rawproduct was recrystallized from a 1-1 acetone-water mixture and afterconcentration, the crystals were recovered by vacuum filtration and werewashed with a 15-50 acetone-water mixture.

2l-m-sulfobenzoate of 6,16a-dimethyl- [3 ,2c]pyrazol- After drying underreduced pressure, 4.05 g (68.5% yield) of 2l-m-sulfobenzoate of6,l6a-dimethyl-2'- phenyl-A -pregnatrieneolo [3 ,2-c ]-pyrazo1- 1lB,17,21-triol-20-one was obtained melting at 280C and having a specificrotation [01],, 103 i 2Cc 1 in ethanol with 50 water). The productoccurred in the form of fine needles soluble in a water-acetone mixtureand a water-ethanol mixture and slightly soluble in acetone, water andehtanol.

Analysis: C H O N S; molecular weight 672.78.

Calculated: %C,66.05; %H,5.99; %N,4.17; %S,4.77.

Found:%C,65.8; %H,6.3: %N,4.0; %S,4.6.

EXAMPLE 2 Sodium salt of 2l-m-sulfobenzoate of 6,16a-dimethyl-2'-phenyl-A -pregnatrieneolo [3,2-c]-pyrazol-l 1,8,17,2l-triol-20-one 1.6g (0.00238 mole) of 2l-m-sulfobenzoate of 6,16- oz-dimethyl-Z-phenyl-A-pregnatrieneolo[3,2-]- pyrazol-l 1B,17,21-triol-20-0ne were added to asolu tion of 23.8 ml of 0.1N sodium hydroxide and 137 ml of distilledwater and the mixture was warmed to obtain a limpid solution and thenthe pH was adjusted to 7.3 by addition of small quantities of the free21-msulfobenzoate of 6,16a-dimethyl-2'-phenyl-A pregnatrieneolo[3,2-c]-pyrazol-l 1,8,17,21-triol- 20-one. The solution was thenfiltered and the filtrate was lyophilized to obtain 1.61 g (97.5% yield)of the sodium salt of 6,16a-dimethyl-2'-phenyl-A pregnatrieneolo[3,2-c]-pyrazol-l 1fi,17,2 l-triol- 20-one in the form of a pale yellowsolid soluble in water.

Analysis C H O N Nas; molecular weight 694.77

Calculated: %C,58; %H,6.14; %N,3.63; %Na,3.0l; %S,4.2.

Found: %C,58.5; %H,6.2; %N,3.8; %Na,3. 1; %S,4.1;

U.V. Spectrum (ethanol):

Max at 228 nm D Max. at 282 nm E Max. at 314 nm E 1.R. Sprectrum(Nujol):

Ketone at 1715""1 aromatic C=C at 1594 and PHARMACEUTICAL COMPOSITIONSA. An injectable preparation consisted of 3 mg of 21-m-sulfobenzoate of6, l 6a-dimethyl'2'-phenyl-A pregnatrieneolo [3,2-c]-pyrazol-1 18,1 7,2l-triol- 20-one and sufficient liquid excipient to make a volume of 2ml.

B. A tablet consisted of 1 mg of the sodium salt of 2 1 m-sulfobenzoateof 6,16a-dimethyl-2-pheny1-A'- pregnatrieneolo 20 one and sufficientexcipient consisting of lactose, amidon, talc and magnesium stearate tomake up the desired tablet weight.

PHARMACOLOGICAL STUDY A. Anti-inflammatory Activity Theanti-inflammatory activity was evaluated in the granuloma with cottontest of Meier et a1. lExperienta. Vol. 6 (1950), p. 469] in which groupsof 8 female rats weighing to g each received an implantation of 2pellets of cotton each weighing 10 mg under the skin of the thorax. Thetest compound was administered subcutaneously in aqueous solutioncontaining [3,2-cl-pyrazol-l lB,17,21-triolsodium hydroxide andpropylene glycol at a dose of 5 or 20 ug/kg in 2 administrations per dayfor 2 days. One group acting as the control received only the vehicle.On the 3rd day, 16 hours after the last injection the is manifested inthe l6the hour and the action is maintained for 7 days with the sameintensity.

C. Lymphopeniant Actitity rats were killed and the pellets with theirgranuloma tis- 5 This test is base on the direct action of corticoids onsue envelope were cut out and weighed fresh and then lymphocytesexpressed by a diminution of the number after drying in an oven at 60Cfor IS hours. The weight of circulation leucocytes. After oneleucocytary countof the granuloma was obtained by substracting theiniing, the 2 l-m-sulfobenzoate was intraperitoneally adtial weight ofthe cotton and the weight of the granuministered to groups of 8 malerats weighing about 100 loma formed in the treated rats was expressed asa per- 10 g in solution in isotonic sodium chloride solute at a centageof the weight of the granuloma in the controls. dose of 200 ug/kg(expressed in molecular equivalent The weight of the thymus removed atthe same time as of dexamethasone). One control group received only thegranulomas permitted an indication of the immedithe vehicle 271624 and48 hours and 7 days after the ate thymolytic activity of the products.The antiadministration, a second counting of leucocytes was inflammatoryactivity of 2l-m-sulfobenzoate of for- I made on the blood taken fromophthalmic plexus. mula l in hich R is hydrogen as compared und the Theleucocytes were counted by a computer after the same conditions withthat of cortivazol (21-acetate of r d bl d c r cl s were hemolyzed withsaponin. .16 -dim thy "p y "-pt g 0 [3, The percentage of diminution ofleucocytes was deterc]-pyrazol-l lB,l7,2l-triol--one as a suspension inan mined by comparison with the controls. The lymisotonic sodiumchloride solute containing 0.2% of pol- 20 phopeniant activity of the2l-msulfobenzoate of the ysorbate 80. The results are reported in TableI. invention was compared to that of cortivazol under the TABLE I sameconditions and the results are reported in Table III.

Dry Granuloma Fresh Thymus 75 TABLE I" Dose 7c of of Administered Weightinhibi- Weight inhibi- Groups in .tg/kg in mg tion in mg tion Percentageof diminution of leucocytes after:

2 7 lo 24 48 7 Controls 0 73.3 309 Hours Hours Hours Hours Hours DaysZl-m-sulfo- 5 45.8 38 305 benzoate 20 38.0 48 Ml 22 2l-m-sulfo-Cortivazol 5 46 l 37 252 19 benzoate 54 52 77 47 5 l 20 31.4 57 192 38Cortivazol l 1 44 33 57 60 0 Table I shows that the 2l-m-sulfobenzoateof the in- Table III shows that the 2lm-sulfobenzoate has a vention hasan anti-inflammatory activity comparable 35 more rapid, more intense andmore durable lymto cortivazol. phopeniant activity than cortivazol andprovokes a strong leucopenia lasting for 7 days while the same ef-Thymolync Activity feet for cortivazol has disappeared. The thymolyticactivity was determined on groups of 6 young male rats weighing about 60g and the test 40 Acute Toxlclty compounds were administered in anisotonic sodium The acute toxicity of 2l-m-sulfobenzoate wasdeterchloride solute intraperitoneally at a dose of 200 ug/kg i d ongmups of f l mi weighing 19 to 23 g (expressed in molecular equivalentof dexamethasone). which had been starved for 6 hours. The said productOne lot serving as the control received only the vehicle. was d i i t dll b ta e ly and intrave- The groups of animals were killed after 48nously in solution in distilled water containing 3% of hours and 7 y oftreatment The thymus was absolute alcohol at doses of 600 and i200ug/kg. The moved and the eight h f s expressed In gvolume administeredin each case was 0.4 ml of 20 g of The Percentage of lhvohlhoh of thethymus of the body weight. After observation for 8 days, nomortalitreated animals was compared to the average value of i were t i dthe thymus weight of the controls. The thymolytic ac- 50 tivity of the21-m-sulfobenzoate was compared to cor- CLINICAL STUDY tivazol,dexamethasone and dexamethasone phosphate C Hi N 1 d under the sameconditions and the results are reporte The Slck person was 56 years Oldand had presented in Table II.

y for 3 years asthmatic crisis of exogemc origin without TABLE hcardiovascular reverberations. Allergological investigation placed inevidence a strong sensibilization to dust 0f involution 0f \hymuSflftetl and streptococci. The patient was treated without suc- Compound8 hours 16 hours 24 hours 48 hours 7 days cess with a combination ofprednisone and antibiotics.

60 Following a Cl'lSlS of paroxystrc dyspnea. the patient receivedintramuscularly 3 mg of the said 2 l -mbenzoate 9 37 43 64 Conivazol 1226 14 36 i5 sulfobenzoate and 15 minutes after the lIljBCllOll. theDexarnethasone 38 41 3 8 dyspnea was controlled. The patient wasinspected the gfiggg z t 34 41 0 next day and did not have any crisisfor 24 hours after 65 the lnjflCIlOIl.

Table ll shows that ZI-m-sulfobenzoate possesses a more durable activitythan cortivasol which difference Case history No. 2

The patient was 47 years old and had asthmatic crisis for 10 yearsaccompanied with an important asthenia and after returning from hisholidays in September, the patient had a strong dyspneic crisis that wasnot halted by oral administration of 10 mg of prednisone. The patientreceived intramuscularly 3 mg of the 21-msulfobenzoate and minutes afterinjection, half hour, the dyspnea was controlled. The patient wasinspected the next day and did not have any crisis for 24 hours afterthe injection.

Case History No. 2

The patient was 47 years old and had asthmatic crisis for 10 yearsaccompanied with an important asthenia and after returning from hisholidays in September, the patient has a strong dyspneic crisis that wasnot halted by oral administration of 10 mg of prednisone. The patientreceived intramuscularly 3 mg of the 21-msulfobenzoate and in one halfhour, the dyspnea was controlled. Moreover, the habitual rhinitisassociated with the patients asthma was also controlled. Theamelioration obtained persisted for 48 hours after the single injection.The patient was treated several times after dyspneic manifestations withan injection of 4 mg of Zl-phosphate of dexamethasone and the effectobtained with this last product was not as prolonged as that obtainedwith 2l-m-sulfobenzoate.

Case History No. 3

The patient was 36 years old and had asthmatic crisis of exogenicorigin. allergological investigation showed a strong sensibilization tocandy, streprococci and vaccine of pasteru Institute CCB (used forbronchical complications of asthma). In September, the patient had astrong dyspneic crisis which could not be controlled with the usualtreatments (theophylline or dexamethasone phosphate). The patientreceived intramuscularly 3 mg of the said 2 l -m-sulfobenzoate andwithin a half hour, the dyspnea and the accompanying hyperemotivitydisappeared and by auscultation, one observed the disappearance of therales. The resulting amelioration was maintained for about 24 hours andthe said results can not be obtained by the administration of2l'phosphate of dexamethasone.

Case History No. 4

The patient, 55 years old, had asthmatic crisis of exogenic originassociated with an allergic rhinitis. Allergological investigationsshowed a strong sensibilization against grass and dust. in September,the patient having a strong dyspneic crisis received instramuscularly 3mg of the 2 l-m-sulfobenzoate and minutes after the injection, thedyspneic crisis was controlled and by auscultation, the rales hadcompletely disappeared. The associated allergic rhinitis alsodisappeared.

.Qass iismlleai.

The patient, 23 years old, had an allergic rhinitis associated withasthma of exogenic origin. Allergological investigation showed a strongsensibility to household dust and to grass pollens (absinthe, armoision,dandelion). The patient who suffered all dyspneic crisis every eveningcould not be controlled after administration of theophyllinesuppositories. The patient then received intramuscularly 3 mg of the2l-m-sulfobenzoate and after a half hour, the dyspnea totallydisappeared and had a strong diminution of the intensity and number ofrales. The amelioration obtained by the administration of the2l-m-sulfobenzoate was maintained for 48 hours. The patient was treatedbefore with the 2lphosphate of dexamethazone verifies that the21-msulfobenzoate effected a more rapid, greater and more durableamelioration.

Case History No. 6

The patient, 14 years old, had eczema and asthmatic crisis of exogenicorigin and allergological investigation showed a strong sensibility todust, feathers, grass pollen and cat fur. Upon return from holidays, thepatient showed a dyspneic crisis every evening received intramuscularly3 mg of the 2l-m-sulfobenzoate and after a half hour, auscultationshowed a strong reduction of rales. Also observed was a diminution ofpruris. On the day after the injection, the patient did not experienceany nocturnal dyspneic crisis.

Case History No. 7 h

The patient, 21 years old, showed asthmatic crisis of exogenic originevolving over several years and allergological investigation showed astrong sensibility to eat fur, to dust, to feathers and to wool. In thecourse of the investigation, the patient showed a strong asthmaticcrisis and received at once 3 mg of the 21-msulfobenzoateintramuscularly. The asthma crisis was calmed in a half hour and alsothe rales and dyspnea disappeared completely. Three days later, newtests on the patient were effected to complete the allergologicalinvestigation releasing a strong dyspneic crisis. The patient receivedintravenously 3 mg of the 2l-m sulfobenzoate. The asthma crisis calmedin 15 minutes and after 30 minutes, auscultation showed a completedisappearance of rales. In the two days after the injection, the patientdid not feel any nocturnal dyspneic crisis.

Case History No. 8

The patient, 65 years old, showed urticaria associated with edema of thetongue and of glottis. Following a strong outburst of urticariaassociated with edema of the eyelids, the tongue and oral mucous, thepatient received intramuscularly 3 mg of the 2l-m-sulfobenzoate and ahalf hour after the injection, a clear reduction of edma of the eyelidsand oral mucous was observed with an attentuation of urticaria spots.After a week, the patient did not notice any edema or urticairia afterthe treatment with the 2l-msulfoben2oate and the action thereof wasrapid and durable.

Case History No. 9

Thee patient. 28 years old, showed asthmatic crisis of exogenic originand allergological investigation showed a sensibility to feathers. dustand diverse microbial germs. After a strong asthmatic dyspnea, thepatient received intramuscularly 3 mg of 2l-m-sulfobenzoate and a halfhour after the administration. a very clear amelioration of the state ofthe patient was observed with a disappearance of dyspnea and a reductionof rales by auscultation. The amerlioration persisted for more than aweek.

Various modifications of the product and process of the invention may bemade without departing from the spirit or scope thereof and it should beunderstood that the invention is to be intended to be limited only asdefined in the appended claims.

I claim:

9 1. An anti-inflammatory composition comprising an effective amount ofa 2l-m-sulfobenzoate of 6,160! dimethyl-2 -phenyl-A -pregnatrieneolo [32 pyrazol-l1B,l7,2l-triol20-ohe of the formula CH3 E JQ wherein R isselected from the group consisting of hydrogen and alkali metal and apharmaceutical carrier. 2. A composition of claim 1 wherein R ishydrogen. 3. A method of treating inflammation in warmblooded animalscomprising administering to warm- LII blooded animals ananti-inflammatorily effective amount of a 2 l-m-sulfobenzoate of6,l6a-dimcthyl-2- phenylA' pregnatrieneolo [3,2-c I-pyrazol- 1 1/3,]7,2l-triol-20-one of the formula if if wherein R is selected from thegroup consisting of hydrogen and alkali metal.

4. The method of claim 3 wherein R is hydrogen.

1. AN ANTI-INFLAMMATORY COMPOSITION COMPRISING AN EFFECTIVE AMOUNT OF A21-M-SULFOBENZOATE OF 6,16A-DIMETHYL-2**1PHENYL-$2,4,6-PREGNATRIENEOLO(3,2-C)-PYRAZOL-11B,1721-TRIOL20-ONE OF THE FORMULA
 2. A composition ofclaim 1 wherein R is hydrogen.
 3. A method of treating inflammation inwarmblooded animals comprising administering to warm-blooded animals ananti-inflammatorily effective amount of a 21-m-sulfobenzoate of 6,16Alpha -dimethyl-2''-phenyl- Delta 2,4,6-pregnatrieneolo(3,2-c)-pyrazol-11 Beta ,17,21-triol-20-one of the formula
 4. The methodof claim 3 wherein R is hydrogen.